Bile acids and their receptors: Potential therapeutic targets in inflammatory bowel disease.

Chronic and recurrent inflammatory disorders of the gastrointestinal tract caused by a complex interplay between genetics and intestinal dysbiosis are called inflammatory bowel disease. As a result of the interaction between the liver and the gut microbiota, bile acids are an atypical class of steroids produced in mammals and traditionally known for their function in food absorption. With the development of genomics and metabolomics, more and more data suggest that the pathophysiological mechanisms of inflammatory bowel disease are regulated by bile acids and their receptors. Bile acids operate as signalling molecules by activating a variety of bile acid receptors that impact intestinal flora, epithelial barrier function, and intestinal immunology. Inflammatory bowel disease can be treated in new ways by using these potential molecules. This paper mainly discusses the increasing function of bile acids and their receptors in inflammatory bowel disease and their prospective therapeutic applications. In addition, we explore bile acid metabolism and the interaction of bile acids and the gut microbiota.

Effect of Cementitious Capillary Crystalline Waterproofing Materials on the Mechanical and Impermeability Properties of Engineered Cementitious Composites with Microscopic Analysis.

Building structures are prone to cracking, leakage, and corrosion under complex loads and harsh marine environments, which seriously affect their durability performance. To design cementitious composites with excellent mechanical and impermeability properties, Engineered Cementitious Composites (ECCs) doped with ultrahigh molecular weight polyethylene short-cut fibers (PE-ECCs) were used as the reference group. Different types (XYPEX-type from Canada, SY1000-type from China) and doses (0%, 0.5%, 1.0%, 1.5%, 2.0%) of Cementitious Capillary Crystalline Waterproofing materials (CCCWs) were incorporated. The effect of CCCWs on the mechanical and impermeability properties of PE-ECCs, and the microscopic changes, were investigated to determine the best type of CCCW to use and the best amount of doping. The results showed that with increasing the CCCW dosage, the effects of both CCCWs on the mechanical and impermeability properties of PE-ECC increased and then decreased, and that the best mechanical and impermeability properties of PE-ECC were achieved when the CCCW dosing was 1.0%. The mechanical properties of the PE-ECC were more obviously improved by XYPEX-type CCCW, with a compressive strength of 53.8 MPa, flexural strength of 11.8 MPa, an ultimate tensile stress of 5.56 MPa, and an ultimate tensile strain of 7.53 MPa, which were 37.95%, 53.25%, 14.17%, and 21.65% higher than those of the reference group, respectively. The effects of the two CCCWs on impermeability were comparable. CCCW-PE-ECC(X1.0%) and CCCW-PE-ECC(S1.0%) showed the smallest permeation heights, 2.6 mm and 2.8 mm, respectively. The chloride ion diffusion coefficients of CCCW-PE-ECC(X1.0%) and CCCW-PE-ECC(S1.0%) exhibited the smallest values, 0.15 × 10-12 m2/s and 0.10 × 10-12 m2/s, respectively. Micromorphological tests showed that the particle size of the XYPEX-type CCCW was finer, and the intensity of the diffraction peaks of C-S-H and CaCO3 of PE-ECC increased after doping with two suitable doping amounts of CCCW. The pore structure was improved, the surface of the matrix was smoother, and the degree of erosion of hydration products on the fiber surface was reduced after chloride ion penetration. XYPEX-type CCCW demonstrated a more obvious improvement in the PE-ECC pore structure.

Polymicrobial Anaerobic Meningitis Detected by Next-Generation Sequencing: Case Report and Review of the Literature.

Background: Anaerobic meningitis is a severe central nervous system infection associated with significant neurological sequelae and high mortality. However, the precise detection of causative pathogen(s) remains difficult because anaerobic bacteria are difficult to culture. Next-generation sequencing is a technology that was developed recently and has been applied in many fields. To the best of our knowledge, the use of next-generation sequencing for cerebrospinal fluid analysis in the diagnosis of anaerobic meningitis has been rarely reported. Case presentation: Here, we report a case of polymicrobial anaerobic meningitis diagnosed using next-generation sequencing of cerebrospinal fluid in a 16-year-old girl. Five species of anaerobic bacteria (Porphyromonas gingivalis, Prevotella enoeca, Campylobacter rectus, Fusobacterium uncleatum, and Actinomyces israelii) were detected by next-generation sequencing and treated with antibacterial agents (ceftriaxone, vancomycin, and metronidazole). The patient responded well to antibacterial treatment. Further inspection revealed bone destruction at the base of the skull, which further confirmed that these bacteria had originated from the oral cavity. One month later, the patient's condition improved significantly. At the same time, we performed a literature review on anaerobic meningitis using studies published in the last 20 years. Conclusions: This case emphasizes the importance of applying metagenomic next-generation sequencing to clinch the clinical diagnosis for patients with central nervous system infection. Metagenomic next-generation sequencing has been reported to be an important diagnostic modality for identifying uncommon pathogens.

Lysosome-targeted ruthenium(II) complexes induce both apoptosis and autophagy in HeLa cells.

Ruthenium complexes with good biological properties have attracted increasing attention in recent decades. In this work, three ruthenium polypyridine complexes containing 5-fluorouracil derivatives as ligands, [Ru(bpy)2(L)]2+ (Ru1), [Ru(phen)2(L)]2+ (Ru2), [Ru(dip)2(L)]2+ (Ru3) (L = 1-((1,10-phenanthroline-5-amino) pentyl)-5-fluorouracil; bpy = 2,2'-bipyridine; phen =1,10-phenanthroline; dip = 4,7-diphenyl-1,10-phenanthroline), were synthesized and characterized. Based on in vitro cytotoxicity tests, Ru3 (IC50 = 7.35 ± 0.39 µM) showed the best anticancer activity among three compounds in the selected cell lines. It is worth noting that Ru3 also exerts less cytotoxicity on LO2 cell lines, with an IC50 value 5 times higher than that on HeLa cells, indicating its selective activity. Mechanism studies revealed that Ru3 can specifically target lysosomes and induce cell apoptosis in a caspase-dependent manner. Specifically, Ru3 can arrest cell cycle at the G0/G1 phase, increase the intracellular reactive oxygen species (ROS) level, and then damage DNA. In short, Ru3 can eventually cause cell death through the synergy of inducing apoptosis and autophagy, which was further proven by western blot assay results.

Emergency health response of Japanese local governments after the “3·11” great earthquake / 上海预防医学

The earthquake occurred in Japan on March 11， 2011 was accompanied by a tsunami， which also led to a leakage accident at the Fukushima Daiichi nuclear power plant. The disaster caused large-scale casualties and property damage. Soma City in Fukushima Prefecture established a disaster response headquarter and held its first meeting 9 minutes after the earthquake. Disaster response can be divided into three stages. In the first phase， life protection is the primary task. In the second phase， the primary tasks are health protection and basic life support. And in the third phase， life quality improvement is the primary task. This article reviewed the public health emergency response of local governments in Japan after the disaster in order to improve the response to public health emergencies in China.

Neutralizing and binding antibody kinetics of COVID-19 patients during hospital and convalescent phases

Knowledge of the host immune response after natural SARS-CoV-2 infection is essential for informing directions of vaccination and epidemiological control strategies against COVID-19. In this study, thirty-four COVID-19 patients were enrolled with 244 serial blood specimens (38.1% after hospital discharge) collected to explore the chronological evolution of neutralizing (NAb), total (TAb), IgM, IgG and IgA antibody in parallel. IgG titers reached a peak later (approximately 35 days postonset) than those of Nab, Ab, IgM and IgA (20[~]25 days postonset). After peaking, IgM levels declined with an estimated average half-life of 10.36 days, which was more rapid than those of IgA (51.25 days) and IgG (177.39 days). Based on these half-life data, we estimate that the median times for IgM, IgA and IgG to become seronegative are 4.59 (IQR 4.12-5.03), 7.78 (IQR 6.71-9.16) and 42.72 (IQR 33.75-47.96) months post disease onset. The relative contribution of IgM to NAb was higher than that of IgG (standardized {beta} regression coefficient: 0.53 vs 0.48), so the rapid decline in NAb may be attributed to the rapid decay of IgM in acute phase. However, the relative contribution of IgG to NAb increased and that of IgM further decreased after 6 weeks postonset. Its assumed that the decline rate of NAb might slow down to the same level as that of IgG over time. This study suggests that SARS-CoV-2 infection induces robust neutralizing and binding antibody responses in patients and that humoral immunity against SARS-CoV-2 acquired by infection may persist for a relatively long time.

Several FDA-approved drugs effectively inhibit SARS-CoV-2 infection in vitro

To identify drugs that are potentially used for the treatment of COVID-19, the potency of 1403 FDA-approved drugs were evaluated using a robust pseudovirus assay and the candidates were further confirmed by authentic SARS-CoV-2 assay. Four compounds, Clomiphene (citrate), Vortioxetine, Vortioxetine (hydrobromide) and Asenapine (hydrochloride), showed potent inhibitory effects in both pseudovirus and authentic virus assay. The combination of Clomiphene (citrate), Vortioxetine and Asenapine (hydrochloride) is much more potent than used alone, with IC50 of 0.34 M.

Virtual reduction and design of 3D printed guide template for comminuted patellar fractures / 中华创伤骨科杂志

Objective:To explore the role of virtual reduction and design of 3D printed guide template in assisting reduction and internal fixation of comminuted patellar fractures.Methods:A retrospective study was conducted of 12 patients who had been treated for closed patellar fractures from March 2016 to April 2019 at Department of Orthopaedics, Jiangxi Provincial People's Hospital. They were 8 males and 4 females with an average age of 35.4 years (from 22 to 51). All their fractures were type 34-C3 (more than 3 fragments) according to the AO/OTA classification. Their preoperative CT data were imported into software Mimics to print a full patella, upper and lower halves of the patella, and a guide template for reduction. During operation, bone fragments were first temporarily fixated with a fine Kirschner wire after combined together with the assistance of reduction templates for upper and lower hemi-patellas. Next, the whole patella was precisely reset and definitely fixated using patellar ring ligation and tension band wire with the assistance of the reduction guide template for full patellar. Recorded were operation time, fluoroscopic frequency, and visual analogue scale (VAS), knee range of motion (ROM) and knee function B?stman scores at the last follow-up.Results:In this group of patients, operation time averaged 90 minutes (from 75 to 120 minutes), fluoroscopic frequency 4.5 times (from 3 to 8 times), follow-up duration 19 weeks (from 16 to 22 weeks), and fracture healing time 14 weeks (from 11 to 17 weeks). At their last follow-up, X-ray showed good patellar morphology, fine internal fixation positions and an average VAS pain scores of 0.3 (from 0 to 1), their knee joints could be fully straightened, their knee ROM averaged 130°(from 100° to 138°), and their knee B?stman scores 28.8 points (from 21 to 30 points), giving 9 excellent and 3 good cases.Conclusion:3D printed reduction guide templates can significantly improve the efficiency and quality of surgical reduction in comminuted patella fractures, enhancing fixation strength and facilitating early and full range exercise of the knee joint to achieve excellent recovery.

Prevalence and control of human diseases caused by Beta Coronavirus / 上海预防医学

Beta coronaviruses (β-CoVs) can infect human and a variety of animals, causing respiratory, intestinal, liver and nervous system diseases of the hosts.Since the outbreak of severe acute respiratory syndrome (SARS) caused by coronavirus in 32 countries and regions in 2002, there have been Middle East respiratory syndrome and human coronavirus HKU1 pneumonia caused by other viruses of this genus.β-CoVs pose a continuous threat to human health with its high transmission efficiency, serious infection consequences and uncertain epidemic outbreaks.Hereby, the epidemic history, virus sources and transmission modes of β-CoVs, as well as the detection and monitoring methods for the virus are reviewed in the paper to provide ideas and measures for better control of human diseases caused by β-CoVs.

Prevalence and control of human diseases caused by Beta Coronavirus / 上海预防医学

Beta coronaviruses (β-CoVs) can infect human and a variety of animals, causing respiratory, intestinal, liver and nervous system diseases of the hosts.Since the outbreak of severe acute respiratory syndrome (SARS) caused by coronavirus in 32 countries and regions in 2002, there have been Middle East respiratory syndrome and human coronavirus HKU1 pneumonia caused by other viruses of this genus.β-CoVs pose a continuous threat to human health with its high transmission efficiency, serious infection consequences and uncertain epidemic outbreaks.Hereby, the epidemic history, virus sources and transmission modes of β-CoVs, as well as the detection and monitoring methods for the virus are reviewed in the paper to provide ideas and measures for better control of human diseases caused by β-CoVs.

Avian influenza viruses (AIVs) H9N2 are in the course of reassorting into novel AIVs / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

In 2013, two episodes of influenza emerged in China and caused worldwide concern. A new H7N9 avian influenza virus (AIV) first appeared in China on February 19, 2013. By August 31, 2013, the virus had spread to ten provinces and two metropolitan cities. Of 134 patients with H7N9 influenza, 45 died. From then on, epidemics emerged sporadically in China and resulted in several victims. On November 30, 2013, a 73-year-old woman presented with an influenza-like illness. She developed multiple organ failure and died 9 d after the onset of disease. A novel reassortant AIV, H10N8, was isolated from a tracheal aspirate specimen that was obtained from the patient 7 d after onset. This case was the first human case of influenza A subtype H10N8. On 4 February, 2014, another death due to H10N8 avian influenza was reported in Jiangxi Province, China.

Hollow hydroxyapatite combined with human bone morphogenetic protein-2 microspheres for bone defect repair / 中国组织工程研究

BACKGROUND:Hydroxyapatite (HA) is a good scaffold material, and recombinant human bone morphogenetic protein-2 (rhBMP-2) possesses a strong osteogenic ability, therefore, by which preparing a novel composite material wil be helpful for bone repair. OBJECTIVE:To explore the effects of the hol ow HA/rhBMP-2 microspheres on the osteogenesis and biomechanics of rabbit bone defects. METHODS:Forty-eight male healthy adult New Zealand white rabbits were randomly divided into three groups (n=16 per group), including composite, single and control groups. Radical defect models were prepared, and the hol ow HA/rhBMP-2 and hol ow HA scaffolds were implanted into the composite and single groups, respectively. The control group received no treatment. At the 1st day of 4, 8, 12, and 16 weeks after implantation, the level of serum alkaline phosphatase was detected, and the bone healing was assessed through X-ray, three-dimensional CT, radionuclide bone scan and biomechanics testing, respectively. RESULTS AND CONCLUSION:The level of serum alkaline phosphatase, X-ray scale scores, osteogetic effect, region of interest volume, three-dimensional CT and biomechanical strength in the composite group were superior to those in the single group. In the meanwhile, the bone healing was unsatisfactory in the control group. Our findings indicate that the hol ow HA/rhBMP-2 artificial bone exhibits a good osteogenic ability and mechanical strength, contributing to bone healing.

Urinary level of tissue factor and its procoagulant activity in patients with type 2 diabetes / 南方医科大学学报

<p><b>OBJECTIVE</b>To examine the urinary level of tissue factor (uTF) and its procoagulant activity (PCA) in patients with diabetes mellitus, and explore the relationship between uTF and renal damage in diabetes mellitus.</p><p><b>METHODS</b>Eighty-six patients with type 2 diabetes mellitus were divided into 3 groups according to urine albumin excretion (UACR), namely normal albuminuria group, microalbuminuria group and macroalbuminuria group. The levels of uTF, PCA, blood urea nitrogen (BUN), serum creatinine (CRE), serum cystatin C (CYSC), glycohemoglobin A1c (HbA1c), and high-sensitivity C-reactive protein (hs-CRP) were measured in all the patients and 21 healthy controls.</p><p><b>RESULTS</b>Compared with normal control, the diabetic patients showed significantly increased levels of uTF and PCA. The urinary TF-PCA was positively correlated to BUN, CYSC, CRE, UACR, fasting glucose and hs-CRP, but not to uTF; only hs-CRP, UACR were positively correlated to uTF.</p><p><b>CONCLUSION</b>uTF is probably implicated in the development and progression of diabetic nephropathy.</p>

Effect of simulated microgravity on human monocytic cell proliferation and tissue factor mRNA expression / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of simulated microgravity on the proliferation of human monocytic cells THP-1 and the expression of tissue factor (TF) mRNA.</p><p><b>METHODS</b>THP-1 cells were cultured under a simulated microgravity environment using the rotating cell culture system (RCCS). The changes in the cell proliferation after microgravity culture were assessed by cell counting and cell cycle analysis with flow cytometry. RT-PCR was used to detect the changes in the expression of TF mRNA in THP-1 cells.</p><p><b>RESULTS</b>Culture under simulated microgravity resulted in a significant decrease in the cell number of THP-1 cells in comparison with that of the control cells (P<0.01). After a 24-h culture under microgravity, the G0-Gl phase cells increased from the control level of (46.57∓1.64)% to (67.64∓2.71)% (P<0.05). The cells in both groups showed a low level of TF mRNA expression in the absence of LPS stimulation. A 4-h stimulation with LPS caused up-regulated expression of TF mRNA in both cells, but the microgravity group showed a significantly smaller increase in the expression (2.301∓0.179) than the control group (9.210∓1.328) (P<0.05).</p><p><b>CONCLUSION</b>Microgravity can inhibit the proliferation of THP-1 cells and suppress the cellular expression of TF mRNA.</p>

Isolation and characterization of human rheumatoid arthritis fibroblast-like synoviocytes / 南方医科大学学报

<p><b>OBJECTIVE</b>To isolate and characterize human rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs).</p><p><b>METHODS</b>The synovial membrane tissues were obtained from 4 RA patients, 1 chondroma patient and 1 healthy subject and FLS were isolated by means of tissue culture. The cell morphology was observed by phase-contrast microscope and the cell surface markers were detected by flow cytometry.</p><p><b>RESULTS</b>The FLSs were successfully cultured from the synovial membrane tissues with good cell homogeneity after the third passage. The FLSs of the 3rd to 7th passages were stable and proliferated actively, followed by slow proliferation and aging since the 8th passage. Flow cytometry showed that the 4th-passage FLSs from the RA patients contained 99.04% CD90(+) cells, 2.73% CD3(+) cells, 0.29% CD3(-)CD19(+) cells, 2.81% CD3(-)CD16(+)CD56(+) cells, 5.89% CD14(+) cells, and 54.17% CD55(+) cells. The presence of interleukin-1 receptor type I (IL-1RI, 158.63-/+20.32 pg/ml) and IL-1beta (4.67-/+0.82 pg/ml) were detected in the cell culture supernatant of the 4th-passage FLSs from the RA patients by enzyme-linked immunosorbent assay ELISA.</p><p><b>CONCLUSION</b>FLSs from RA patients can be effectively culture by means of tissue culture, and the cultured FLSs show high expressions of CD90, IL-1RI and IL-1beta.</p>

Inhibition of tissue factor expression in endothelial cells by lentivirus mediated shRNA / 中南大学学报(医学版)

OBJECTIVE@#To construct the recombinant lentivirus RNAi vector, and to determine whether the lentivirus mediated short hairpin RNA (shRNA) can inhibit the tissue factor (TF) expression in endothelial cells.@*METHODS@#Two short hairpin RNAs targeting to human TF were cloned into pENTRTM/U6 plasmid to obtain an entry clone, and the positive clones were verified by sequencing. A recombination reaction was performed between the pENTR/U6 entry construction and pLenti6/BLOCKiTTM-DEST vector, and then the positive clones were confirmed by sequencing. The 293FT cell line was transfected by the above recombined plasmid and lentivirus packing materials, the culture supernatant was harvested, and the virus titer was determined. RT-PCR and ELISA were used to observe the inhibition of TF gene expression after the lentivirus transduction in human umbilical vein endothelial cells.@*RESULTS@#The shRNA sequences targeting to human TF were cloned into the vectors, and an entry clone and an expression clone were constructed successfully, which were proved by sequence determination. Viral particles were packaged in the 293FT cell line, all virus stocks were collected, and the transfection titer was 5*10(5)/transduced unit. RT-PCR and enzyme linked immunosorbent assay demonstrated that the lentivirus stocks could suppress the TF expression in endothelial cells remarkably.@*CONCLUSION@#Lentivirus RNAi vectors containing human TF gene are successfully constructed, and lentivirus mediated shRNA can inhibit the TF expression in endothelial cells, which may provide a highly effective method for the prevention and treatment of thrombo-embolic diseases.

An investigation on hepatitis E virus infection status among livestock in Xi'an area / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the carrier state of hepatitis E virus(HEV) in livestock in Xi'an area.</p><p><b>METHODS</b>Bile samples from swine, canine, sheep and cow were collected from a local slaughtering house. Reverse transcriptase nested polymerase chain reaction (RT-nPCR) was employed to amplify the ORF2 region in HEV RNA genome. All positive samples were sequenced and compared with data from GenBank. Homology analysis was conducted based on the outcome of sequencing.</p><p><b>RESULTS</b>194, 178, 79 and 191 bile samples from swine, canine, cow and sheep were collected. Positive rates with RT-nPCR method in these domestic animals were 4.10%, 0%, 0% and 0% respectively. Genetic distance analysis indicated that strains being identified were close to genotype IV of HEV, then genotype I, II and III in nucleic acid. Same outcome was shown by the same analysis on amino acid.</p><p><b>CONCLUSION</b>Swine was the only reservoir of HEV in livestock and genotype IV was the prevalent genotype.</p>

Value of plasma tissue factor, tissue factor pathway inhibitor and factor VII assessments in patients with acute myocardial and cerebral infarction / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the clinical implications of changes in plasma tissue factor (TF), tissue factor pathway inhibitor (TFPI) and factor VII (FVII) after the onset of acute myocardial infarction (AMI) and acute cerebral infarction (ACI).</p><p><b>METHODS</b>Sixty-nine patients with AMI, 71 with ACI and 50 age-matched healthy volunteers were enrolled in this study. Blood samples were obtained from the healthy subjects and from the patients at the early stage of AMI and ACI onset for examination of plasma TF and TFPI activity using chromogenic assay, and the plasma TF and TFPI antigens were measured by enzyme-linked immunosorbent assay (ELISA). The plasma FVII coagulation activity (FVII:C) was also measured, and the plasma FVIIa determined using soluble TF assay.</p><p><b>RESULTS</b>Compared with the healthy control group, AMI patients had significantly enhanced plasma TF and TFPI activities and elevated TF and TFPI antigen levels (P<0.05), with also markedly increased FVIIa (P<0.05) but comparable FVII:C (P>0.05). In ACI patients, the plasma TF activity and antigen were obviously increased in comparison with the control group (P<0.05), but plasma TFPI activity and antigen were lowered (P<0.05), and both the FVII:C and FVIIa were markedly higher (P<0.05). Significant differences were noted in plasma TF and TFPI activities and their antigen levels as well as in FVII:C, but not in FVIIa between AMI and ACI patients.</p><p><b>CONCLUSION</b>V Following the onset of AMI and ACI, TF pathway is initiated and the risk of thrombogenesis increases, and the assessment of TF pathway is therefore of value for understanding the development of the condition.</p>

Molecule character of rabies virus in 3 counties in Hunan China / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the rabies molecular biology features in animals between high incidence area of rabies and no rabies cases area in Hunan.</p><p><b>METHODS</b>detect saliva of dogs and brains of dogs and cats by direct immunofluorescence assay, review positive samples by RT-PCR, sequencing extract RNA virus for genetic analysis.</p><p><b>RESULTS</b>12 were detected rabies virus antigen and positive nucleoside acid in 82 dogs from Wugang city also 1 in 17 from Dongkou county; the positive rate: Wugang 14.63 percent, Dongkou 5.88 percent. No rabies virus was detected in 67 samples of dog brains from Fenghuang County. Also none in 28 samples of cat brains. Amplificating N gene of rabies virus from positive samples of dog brain's tissue (No Wg13, Dk13) by RT-PCR, it shows that homology of nucleoside acid between two strain of virus is 99.4 percent; also 99.1 percent of amino acid. The homology of nucleoside acid (amonio acid) among Wg13 stain and Chinese strain CTN and aG strain is 89.4 percent (98.2 percent) and 86.1 percent (95.1 percent); The homology of nucleoside acid (amonio acid) among Dk13 stain Chinese strain CTN and aG strain is 89.1 percent (98.0 percent), 86.1 percent (94.9 percent).Compare with isolated rabies virus from abroad, the homology between two strains and Indonesia is 92.8 percent and 93.2 percent, the most similar of them. The strains isolated from other countries including Japan, Sri Lanka and India are relatively lower; The sequence of gene Wg13 and Dk13 were taken replacement of amino acid.</p><p><b>CONCLUSION</b>Two strains are belong to type I rabies virus, comparing its N gene with current using vaccine strains, both are in same group, and homology are relatively higher.</p>

Analyses on the isolation and characterization of canine rabies virus isolates in Henan / 中华流行病学杂志

<p><b>OBJECTIVE</b>To analyze the genetic characteristics of two rabies virus isolates from Henan province and to compare their relations hip with known rabies virus isolates and vaccine strains.</p><p><b>METHODS</b>Rabies viral antigens were detected in 100 canine brains by immunofluorescence assay method. Rabies virus was isolated through inoculating the suspensions of positive brains into suckling mice. N gene and G were amplified by RT-PCR and sequenced. Phylogenetic trees were constructed for the analysis on genetic characteristics of rabies virus.</p><p><b>RESULTS</b>Two rabies virus strains were isolated (Henan Hb1 and Henan Sq1). Data from sequential comparison revealed that the nucleotide and amino acid identities of N and G gene between the two isolates were 99.3% and 98.9%, and 98.7% and 98.4% respectively. The two isolates were more closely related to CTN, with the homogeny of N gene and G gene as 89.1% and 85.6%-85.7% at the nucleotide level, but 97.6%-98.0% and 92.3% at the amino acid level respectively, than to other vaccine strains. When comparing with other known viruses including Chinese isolates, the two stains shared closer identity with the isolates from Indonesia, and the rates of homogeny of N and G gene were 92.1%-93.2% and 91.9%-92.1% at the nucleotide level, 97.5%-98.6% and 96.0%-96.2% at the amino acid level, respectively. Data from the deduced amino acid sequences revealed that some amino acid residues including the residues in the N and G antigenic sites were substituted in the two isolates. Furthermore, phylogenetic analysis showed that the two isolates were also more closely related to the strains from Indonesia and vaccine strain CTN than to any other known street viruses and vaccine strains.</p><p><b>CONCLUSION</b>Both Henan Hbl and Henan Sql belonged to genotype 1. However, the N and G gene diverged from known street viruses and vaccine strains at either nucleotide level or amino acid level.</p>